Published online before print April 17, 2008, doi: 10.1161/ATVBAHA.108.166405
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© 2008 American Heart Association, Inc.
Integrative Physiology/Experimental Medicine |
Role of Smooth Muscle cGMP/cGKI Signaling in Murine Vascular Restenosis
From the Institut für Pharmakologie und Toxikologie (R.L., P.W., D.B., F.H.), TU München, Germany; Interfakultäres Institut für Biochemie (S.F., R.F.), Universität Tübingen, Germany; Max-Delbrück-Centrum für Molekulare Medizin (M.G.), Berlin-Buch, Germany; Department of Molecular Genetics (J.H.), UT Southwestern, Dallas, Tex; Deutsches Herzzentrum (S.M.), TU München, Germany; and Institut für Kardiovaskuläre Molekularbiologie (A.Z., C.W.), RWTH Aachen, Germany.
Correspondence to Dr Robert Lukowski, Institut für Pharmakologie und Toxikologie der TUM, Biedersteiner Str. 29, D-80802 München, Germany. E-mail lukowski{at}ipt.med.tu-muenchen.de
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Abstract |
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Background— Nitric oxide (NO) is of crucial importance for smooth muscle cell (SMC) function and exerts numerous, and sometimes opposing, effects on vascular restenosis. Although cGMP-dependent protein kinase type I (cGKI) is a principal effector of NO, the molecular pathway of vascular NO signaling in restenosis is unclear. The purpose of this study was to examine the functional role of the smooth muscle cGMP/cGKI signaling cascade in restenosis of vessels.
Methods and Results— Tissue-specific mouse mutants were generated in which the cGKI protein was ablated in SMCs. We investigated whether the absence of cGKI in SMCs would affect vascular remodeling after carotid ligation or removal of the endothelium. No differences were detected between the tissue-specific cGKI mutants and control mice at different time points after vascular injury on a normolipidemic or apoE-deficient background. In line with these results, chronic drug treatment of injured control mice with the phosphodiesterase-5 inhibitor sildenafil elevated cGMP levels but had no influence on the ligation-induced remodeling.
Conclusions— The genetic and pharmacological manipulation of the cGMP/cGKI signaling indicates that this pathway is not involved in the protective effects of NO, suggesting that NO affects vascular remodeling during restenosis via alternative mechanisms.
Key Words: nitric oxide • PKG • atherosclerosis • carotid ligation • wire-injury
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Introduction |
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Occlusion of coronary arteries results from advanced atherosclerotic lesions or plaque rupture.1 A common therapy to restore the blood flow in such narrowed arteries is angioplasty. However, endovascular interventions may lead to a long-term risk of restenosis, ie, the remodeling and eventually reocclusion of treated arteries. Although the exact pathogenesis of restenosis is unclear, it is known that after the initial injury smooth muscle cells (SMCs) contribute together with other cell types to the remodeling of the vessel wall.
See accompanying article on page 1207
Nitric oxide (NO) is an important regulator of cardiovascular homeostasis and, in particular, of SMC function.2 In addition to potent vasorelaxant properties of NO, its bioavailability and signaling have also been associated with vascular proliferative diseases, such as atherosclerosis and restenosis.3–6 Studies with transgenic mice that overexpress or lack the NO synthases indicated that NO can mediate beneficial effects on the vasculature under particular pathophysiological conditions.7–11 On the other hand, several studies reported that NO promoted the progression of vasculoproliferative processes, and, thus, that it was deleterious for the vasculature.12–15 In restenosis, the endogenous effector(s) and underlying molecular mechanism(s) that mediate the opposing effects of NO on vascular remodeling are not clear. Under physiological conditions, NO exerts many of its effects via stimulation of the soluble guanylyl cyclase (sGC), which generates the second messenger cyclic guanosine-3'-5'-monophosphat (cGMP). The cGMP-dependent protein kinase type I (cGKI) is presumably the major target of NO/cGMP signaling in SMCs.2 For example, mice with a homozygous deletion of the cGKI gene showed an impaired NO/cGMP-dependent relaxation of SMCs.16 The role of cGKI has also been implicated in the phenotypic modulation of SMCs, which occurs in atherosclerosis and restenosis.17,18 Several studies suggested that activation of cGKI-dependent pathways has antimitogenic effects in SMCs in vitro,19–21 and might be vasculo-protective in vivo.22,23 Recent evidence suggested that elevation of cGMP by the phosphodiesterase-5 (PDE-5) inhibitor sildenafil, hence, activation of cGMP-dependent pathways, was beneficial for the treatment of hypertrophic heart disease24 and for vascular remodeling associated with pulmonary hypertension.25 These findings indicated that particularly activation of cGKI mediated the protective effects of sildenafil in vivo.26
Surprisingly, the analysis of cGKI-deficient mice produced some evidence that activation of cGKI in SMCs increased the growth of primary SMCs and promoted the formation of atherosclerotic lesions in the apolipoprotein E (apoE)-deficient mouse model of hyperlipidemia-induced atherosclerosis.27 These results supported a unique proatherogenic potential of SMC cGKI in atherosclerosis,18 whereas the direct role of endogenous cGMP/cGKI signaling in restenosis, to our knowledge, was not tested before.
In the present study, we addressed the question whether the cGMP/cGKI pathway in SMCs is also involved in the vascular pathology of restenosis. The Cre/loxP system was used to generate mice with a tissue-specific deletion of the cGKI gene in SMCs. To test the pathophysiological role of endogenous cGKI 2 established mouse models, which resemble restenosis in humans, were used: the unilateral cessation of blood flow by carotid ligation28 and the removal of the endothelium by wire-injury of the carotid artery.29
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Materials and Methods |
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Detailed methods are described in the supplemental data (available online at http://atvb.ahajournals.org).
Experimental Animals
The SM22
-Cre mouse line30 was crossed to mice that carried loxP-flanked cGKI alleles (L2)31 to generate cGKI mutants with recombined cGKI-null alleles (L-). The tissue-specific knockout of the cGKI gene was generated in cardiac and smooth muscle cells (cGKIcsmko, genotype: SM22-Cretg/+; cGKIL-/L2). For experiments theses cGKIcsmko animals were compared to littermate controls (ctr, genotype: SM22-Cretg/+; cGKI+/L2). All animals were maintained and bred in the animal facility of the Institut für Pharmakologie und Toxikologie, Technische Universität München and had access to water and standard chow ad libitum. All experimental procedures were conducted according to the local government’s committee on animal care and welfare in München.
Carotid Artery Ligation
The injury procedure was performed by adapting an established model.28 Briefly, the left common carotid artery of deeply anesthetized animals was dissected and completely ligated proximal to its bifurcation.
Endothelial Denudation
Surgery was performed as described previously.29 Endothelial denudation of the left common carotid artery was performed by withdrawal injury passing a 0.014 inch flexible angioplasty guide wire 3 times trough the vessel. This method very efficiently removed the endothelium.
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Results |
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Tissue-Specific Deletion of cGKI in SMCs
The recombination properties, in particular the tissue-specificity, of the SM22
-Cre mouse line (SM22-Cre)30 were evaluated by crossing Cre transgenic animals to ROSA26 Cre reporter (R26R) mice.32 X-Gal staining of tissues from SM22-Cre; R26R reporter mice for β-galactosidase activity demonstrated recombination in the heart (Figure 1a) and in the common carotid artery (CCA; Figure 1b), whereas with the exception of cardiomyocytes, no staining could be detected in all non-SMC tissues analyzed (data not shown). At the cellular level, blue staining was visible in vascular SMCs of the medial cell layer of the uninjured CCA (Figure 1c). In addition, recombination in the SM22-Cre; R26R reporter mice was analyzed 28 days after carotid ligation. At this time point, severe morphological changes were apparent in the ligated vessel. Inside the internal elastical lamina (IEL), a prominent neointima (NI) had formed. As shown by X-gal staining of ligated vessels, recombination was detectable in the media and importantly became manifest in cells of the NI as well (Figure 1d). These results demonstrated that Cre activity in the SM22-Cre mouse line is restricted to cardiomyocytes and SMCs and that the recombination efficiency in medial SMCs of the CCA as well as in NI forming cells after injury is very high.
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A cGKI-null allele (L-) in SMCs was generated by crossing the SM22-Cre mouse line to mice carrying a conditional loxP-flanked cGKI allele (L2).31 Cre-mediated recombination of the loxP sites and the resulting L- allele was detectable by polymerase chain reaction (PCR) analysis of genomic DNA in SMC-containing tissues and in the heart (supplemental Figure I). Importantly, recombination of the L2 to the L- allele was highly efficient in the CCA and the aorta, less efficient in heart, and not detectable in non–smooth muscle tissue. Western blot analysis demonstrated that the level of cGKI protein was strongly reduced in the CCA and aorta, moderately reduced in the heart, and not altered in the cerebellum (Figure 2a). The decrease in cGKI protein levels in the hearts of the cGKIcsmko mice might result from both a transient SM22 promotor fragment activity and subsequent Cre expression in the embryonic cardiomyocyte lineage33 as well as from a persistent expression of the Cre transgene in coronary SMCs. In uninjured control animals, immunohistochemical analysis of the CCA (Figure 2b) and aorta (data not shown) using specific antibodies34 revealed a strong and uniform expression of the cGKI protein in the medial layer. At 28 days after ligation of the left CCA from control animals, the cGKI protein was expressed in the media and, in addition, it was detectable in cells of the neointima (NI) (Figure 2b). However, the heterogeneous expression pattern observed in the NI probably resulted from both a restricted downregulation of cGKI in some media derived SMCs that switched from the contractile to a more synthetic/proliferative phenotype,17 and the contribution of cGKI-negative non-SMCs to the NI formation. In cGKIcsmko mutants, no cGKI staining could be identified in the uninjured CCA and 28 days after carotid ligation (Figure 2b). Thus, the genetic strategy via Cre/loxP-mediated somatic mutagenesis resulted in an efficient ablation of the cGKI protein from the uninjured and injured CCA. However, under normal conditions the cGKIcsmko mice were healthy and showed no gross phenotypic abnormalities as demonstrated by their long term survival rates and a normal gastrointestinal transit time (supplemental Figure II). Importantly, the arterial morphology and vessel structure as analyzed by the expression of several cGKI substrates and SMC marker proteins showed no significant differences between both genotypes before vascular injury (supplemental Figure III). In addition, the basal levels of endogenous cAMP (supplemental Figure IVa) and cGMP (supplemental Figure IVb) of control and cGKIcsmko vascular SMCs were similar. In both genotypes treatment of vascular SMCs with sildenafil (0.1 mmol/L) or the NO-donor DEA-NO (0.1 mmol/L) increased cGMP-levels by 
20 and 80-fold respectively, whereas cAMP levels showed no significant changes (supplemental Figure IV). Thus, the genetic approach to abolish cGKI activity in vascular SMCs did not affect endogenous cyclic nucleotide levels. In addition, the 8-Br-cAMP–stimulated phosphorylation of VASP at Ser157, a common phosphorylation site for both cGKI- and cAMP-dependent kinases (cAKs), was detectable in primary vascular SMCs of control as well as cGKIcsmko mice (Figure 3). However, the 8-Br-cGMP–stimulated phosphorylation of substrate proteins was clearly impaired in vascular SMCs of the cGKIcsmko as demonstrated by the use of phospho-specific antibodies (Figure 3). These results support the conclusion that vascular SMCs of tissue-specific cGKIcsmko mice lack cGKI activity.
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Vascular Remodeling After Carotid Ligation
Ligation of the left CCA induced intensive remodeling of the injured vessel segment in both control (genotype: SM22-Cretg/+; cGKI+/L2) and cGKIcsmko (genotype: SM22-Cretg/+; cGKIL-/L2) mice. The histological analysis of hematoxylin and eosin (H&E)-stained sections showed the formation of a neointima within the IEL, but no obvious differences were found between the genotypes 28 days after injury (supplemental Figure Va). The presence of an intact endothelial cell layer on the luminal surface of the ligated vessels was demonstrated by immunoreactivity for the von Willebrand factor (vWF) at all time-points analyzed (data not shown). The positive vWF stain confirmed the existence of endothelial cells after injury, and, therefore, it was anticipated that the endothelial sources of NO were preserved during the entire injury period. Furthermore, by studying knockout mice it was shown before that loss of NO derived from the NO synthase (NOS) isozymes had a major impact on the remodeling process in the ligation model.9,35
Quantitative assessment of the remodeling response was performed by morphometric analysis on H&E stained sections. Based on the mean NI/media ratios, no differences could be detected between control mice (NI/media ratio 0.38±0.06; n=22) and littermate cGKIcsmko mice (NI/media ratio 0.45±0.08; n=10) 28 days after injury (Figure 4a). As expected, the extent of remodeling was less intensive after 14 days, but again no differences could be detected between both groups (ctr NI/media ratio 0.12±0.03; n=18; cGKIcsmko NI/media ratio 0.14±0.04; n=13; Figure 4a). To test whether an activation of the cGMP/cGKI pathway after injury had consequences for the vascular response to ligation, a control group of animals (genotype: SM22-Cretg/+; cGKI+/L2) was continuously treated with sildenafil in their drinking water. As determined by the NI/media ratio 28 days after injury, no significant difference was detected between animals treated with sildenafil (NI/media ratio 0.29±0.10; n=10) and the untreated control mice (Figure 4a), although cGMP levels were effectively elevated in the drug-treated mice (supplemental Figure Vb).
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To focus on possible local consequences of the cGKI inactivation, continuous NI/media profiles of the injured arteries were generated (Figure 4b). These profiles revealed a gradual decline of the NI/media ratio with increasing distance to the point of ligation and confirmed a similar response to injury of control and cGKIcsmko animals. Additional vessel parameters, such as the area inside the external elastic lamina (EEL) (ctr 58.1±4.8x103µm2; n=22; cGKIcsmko 72.8±8.1x103µm2; n=10), the medial area (ctr 25.8±1.6x103µm2; n=22; cGKIcsmko 31.3±2.9x103µm2; n=10), the NI area (ctr 10.8±1.8x103µm2; n=22; cGKIcsmko 14.5±2.7x103µm2; n=10), and the area of the vessel lumen (ctr 21.6±2.3x103µm2; n=22; cGKIcsmko 26.9±3.9x103µm2; n=10) again demonstrated that controls and cGKIcsmko responded equally to the injury (Figure 4c). All vascular parameters were also determined after 14 days of injury, but again they were similar for both genotypes (data not shown).
As shown previously, proliferation of cells from the vessel wall was involved in the remodeling after vascular injury.28 To assess this issue, the density of a proliferation marker was evaluated in the remodeled vessel (supplemental Figure Vc). The percentage of positive stained cells in the NI and the medial cell layer was comparable between genotypes. This result indicated that proliferation of cells from the vessel wall contributed to remodeling, but the deletion of cGKI in SMCs showed again no influence.
Earlier reports suggested that SMC cGKI modulated the progress of atherogenesis.27 It was possible that a contribution of cGKI to vascular remodeling after mechanical injury required factors that are present in atherosclerosis-prone mice. Therefore, we reinvestigated vascular remodeling in apoE-deficient (apoE-ko) mice using the above model of CCA ligation. The morphometric data based on measurements of the NI/media ratio (apoE-ko ctr NI/media ratio 0.59±0.09; n=13 and littermate apoE-ko cGKIcsmko NI/media ratio 0.79±0.15; n=12; Figure 5a) and all other vessel parameters such as the EEL (apoE-ko ctr 86.8±6.6x103 µm2; n=13; apoE-ko cGKIcsmko 94.1±10.9x103 µm2; n=12), the medial area (apoE-ko ctr 36.7±2.8x103 µm2; n=13; apoE-ko cGKIcsmko 37.5±4.3x103 µm2; n=12), the NI (apoE-ko ctr 21.3±3.2x103 µm2; n=13; apoE-ko cGKIcsmko 30.9±7.4x103 µm2; n=12), and the lumen (apoE-ko ctr 28.7±2.6x103 µm2; n=13; apoE-ko cGKIcsmko 25.7±3.7x103 µm2; n=12) revealed that both genotypes responded equally to the ligation (Figure 5b). As shown before in the NI and media of normolipedemic mice, the cGKI protein was expressed 28 days after injury in apoE-deficient control mice, and this expression pattern was abolished in apoE-deficient cGKIcsmko mutants (supplemental Figure VI). Thus, the comparable remodeling responses of apoE-deficient control and cGKIcsmko mice did not result from an inadequate recombination efficiency of the SM22-Cre transgene because vascular cGKI was absent in the mutants. We cannot exclude that the lack of cGKI during ontogenesis in the present injury model perhaps led to an undetected functional compensation of the pathway, whereas in the atherosclerosis model, a tamoxifen-inducible Cre was used to delete cGKI selectively only in SMCs of adult mice that were fed an atherogenic diet.27
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Vascular Remodeling After Wire-Injury of the CCA
All experiments shown so far used ligation of the left CCA. It was therefore hypothesized that the inability to detect an effect of SMC cGKI on vascular remodeling was perhaps caused by the presence of an "intact" endothelium. We therefore removed the endothelium by the wire-injury procedure.29 In line with the abolished cGKI expression pattern observed in ligated vessels of cGKIcsmko mutants, no cGKI protein was expressed in the cGKIcsmko 28 days after wire-injury (supplemental Figure VII). The data based on a detailed analysis of the NI/media ratio profile (Figure 6a) and all morphometric parameters of the remodeled vessel (Figure 6b) revealed no difference between both genotypes. In the vessel segment analyzed, the areas of the EEL (ctr 63.9±6.6x103 µm2; n=9; cGKIcsmko 64.6±4.3x103 µm2; n=11), the media (ctr 23.4±1.3x103 µm2; n=9; cGKIcsmko 25.3±1.8x103 µm2; n=11), the NI (ctr 33.1±6.0x103 µm2; n=9; cGKIcsmko 28.6±3.2x103 µm2; n=11), and the lumen (ctr 6.6±1.0x103 µm2; n=9; cGKIcsmko 10.7±1.7x103 µm2; n=11) were similar between genotypes after denudation. However, in a small region at approximately 1.4 to 1.8 mm distance from the bifurcation, the detailed analysis of the profiles revealed a restricted but statistically significant decrease in the NI/media ratios of the cGKIcsmko mutant mice.
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Discussion |
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Recent analysis of vascular proliferative disorders in cGKI mouse mutants indicated that smooth muscle cGKI accelerates plaque formation in a hyperlipidemia-induced model of atherosclerosis.27 These results suggested that cGKI in SMCs potentially mediates proatherogenic properties of the NO/cGMP pathway in vivo, being thus, in contrast to the common view, not vasculoprotective. To solve the controversy whether the cGMP/cGKI pathway in SMCs accelerates lesion formation or mediates vascular protection, we generated different models of vascular restenosis. In comparison to control mice, we could not detect any influences of the cGKI deletion in SMCs on the NI/media ratio, proliferation, and various other vessel parameters at different time-points after injury in normolipedemic and in apoE-deficient mice. In line with these results, activation of the cGMP/cGKI pathway by chronic administration of sildenafil had no influence on the remodeling response after carotid ligation. Further, no potentially vasculoprotective effects of endogenous cGKI on the remodeling were observed in the absence of a functional endothelium. Interestingly, the tissue-specific deletion of cGKI in SMCs had a minor beneficial impact after wire-injury in a spatially restricted vessel segment. The inability of smooth muscle cGKI to affect remodeling was not caused by an altered distribution of the enzyme between media and NI. Previous analysis of the NI formation after balloon injury of rat carotid arteries showed that the media and the NI express cGKI after injury.36 These results implied a potential role of the cGMP/cGKI pathway for injury-induced remodeling, and identical data have been obtained in the present study.
Neither genetic nor pharmacological manipulation of cGMP/cGKI signaling indicate that smooth muscle cGKI is of major importance during mechanically-induced restenosis of murine vessels. It has been suggested that the effects of NO are strongly concentration-dependent. Presumably, the spatio-temporal profile, the amount of NO synthesized, and the source of its production after vascular injury result in the activation of alternative mechanisms that are not mediated by cGKI or cGMP, such as redox regulation of target proteins.37,38 It has been reported recently that cGMP-independent redox regulation of the sarcoplasmatic reticulum calcium ATPase (SERCA) might be required for NO inhibition of SMC migration after arterial injury.39
The cGMP/cGKI pathway has been previously evaluated in models that differ from the ligation model. In rat the balloon injury model has been used to examine the contribution of cGMP and cGKI to the restenosis process.23,40,41 Adenoviral transfer of a constitutively active kinase domain of cGKI23 and sGC41 in conjunction with the application of the NO-donor molsidomine reduced significantly the proliferation of cells in the NI. In contrast, adenoviral overexpression of the full-length cGKIβ isoform did not affect restenosis induced by balloon injury.23 In the same model, YC-1, an activator of sGC, prevented NI formation.40 In these studies endogenous cGMP levels after drug treatment were not determined, and it was not clear whether or not the observed effects were cGMP-dependent. Interestingly, it was shown recently that YC-1 activates matrix metalloproteinases thereby inhibiting neointima formation in a cGMP-independent manner.22 Together, the pharmacological and genetic approach used in the present work differs significantly from balloon injury in conjunction with adenoviral gene transfer.
In conclusion, the present study demonstrated that neither a deletion of cGKI in SMCs nor an activation of the cGMP/cGKI pathway through pharmacological PDE-5 inhibition impaired the overall vascular remodeling after carotid ligation in normolipidemic and apoE-deficient or wire-injured mice. All experimental approaches tested the influence of endogenous SMC cGKI on vascular remodeling by locally restricted injuries resembling restenosis, whereas the analysis of atherosclerosis identified a proatherogenic role of SMC cGKI for systemic vessel disease.27 Based on the presented results, we conclude that the well-accepted effects of NO in restenosis35 are independent of smooth muscle cGKI and that the contribution of cGKI-mediated mechanisms to vascular remodeling appears to be context-specific, being more important in atherosclerosis than in restenosis.
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Acknowledgments |
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We are grateful to Sabine Brummer for excellent technical help.
Sources of Funding
This work was supported by grants from the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie. R.L. and D.B. were fellows of the GRK333 and GRK438, respectively.
Disclosures
None.
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Footnotes |
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Original received October 29, 2007; final version accepted April 4, 2008.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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1. Ross R. Atherosclerosis–an inflammatory disease. N Engl J Med. 1999; 340: 115–126.
2. Feil R, Lohmann SM, de Jonge H, Walter U, Hofmann F. Cyclic GMP-dependent protein kinases and the cardiovascular system: insights from genetically modified mice. Circ Res. 2003; 93: 907–916.
3. Varenne O, Pislaru S, Gillijns H, Van Pelt N, Gerard RD, Zoldhelyi P, Van de Werf F, Collen D, Janssens SP. Local adenovirus-mediated transfer of human endothelial nitric oxide synthase reduces luminal narrowing after coronary angioplasty in pigs. Circulation. 1998; 98: 919–926.
4. von der Leyen HE, Gibbons GH, Morishita R, Lewis NP, Zhang L, Nakajima M, Kaneda Y, Cooke JP, Dzau VJ. Gene therapy inhibiting neointimal vascular lesion: in vivo transfer of endothelial cell nitric oxide synthase gene. Proc Natl Acad Sci USA. 1995; 92: 1137–1141.
5. von der Thusen JH, Fekkes ML, Passier R, van Zonneveld AJ, Mainfroid V, van Berkel TJ, Biessen EA. Adenoviral transfer of endothelial nitric oxide synthase attenuates lesion formation in a novel murine model of postangioplasty restenosis. Arterioscler Thromb Vasc Biol. 2004; 24: 357–362.
6. Janssens S, Flaherty D, Nong Z, Varenne O, van Pelt N, Haustermans C, Zoldhelyi P, Gerard R, Collen D. Human endothelial nitric oxide synthase gene transfer inhibits vascular smooth muscle cell proliferation and neointima formation after balloon injury in rats. Circulation. 1998; 97: 1274–1281.
7. Kawashima S, Yamashita T, Ozaki M, Ohashi Y, Azumi H, Inoue N, Hirata K, Hayashi Y, Itoh H, Yokoyama M. Endothelial NO synthase overexpression inhibits lesion formation in mouse model of vascular remodeling. Arterioscler Thromb Vasc Biol. 2001; 21: 201–207.
8. Moroi M, Zhang L, Yasuda T, Virmani R, Gold HK, Fishman MC, Huang PL. Interaction of genetic deficiency of endothelial nitric oxide, gender, and pregnancy in vascular response to injury in mice. J Clin Invest. 1998; 101: 1225–1232.[Medline] [Order article via Infotrieve]
9. Morishita T, Tsutsui M, Shimokawa H, Horiuchi M, Tanimoto A, Suda O, Tasaki H, Huang PL, Sasaguri Y, Yanagihara N, Nakashima Y. Vasculoprotective roles of neuronal nitric oxide synthase. Faseb J. 2002; 16: 1994–1996.
10. Rudic RD, Shesely EG, Maeda N, Smithies O, Segal SS, Sessa WC. Direct evidence for the importance of endothelium-derived nitric oxide in vascular remodeling. J Clin Invest. 1998; 101: 731–736.[Medline] [Order article via Infotrieve]
11. Kuhlencordt PJ, Gyurko R, Han F, Scherrer-Crosbie M, Aretz TH, Hajjar R, Picard MH, Huang PL. Accelerated atherosclerosis, aortic aneurysm formation, and ischemic heart disease in apolipoprotein E/endothelial nitric oxide synthase double-knockout mice. Circulation. 2001; 104: 448–454.
12. Kuhlencordt PJ, Chen J, Han F, Astern J, Huang PL. Genetic deficiency of inducible nitric oxide synthase reduces atherosclerosis and lowers plasma lipid peroxides in apolipoprotein E-knockout mice. Circulation. 2001; 103: 3099–3104.
13. Ozaki M, Kawashima S, Yamashita T, Hirase T, Namiki M, Inoue N, Hirata K, Yasui H, Sakurai H, Yoshida Y, Masada M, Yokoyama M. Overexpression of endothelial nitric oxide synthase accelerates atherosclerotic lesion formation in apoE-deficient mice. J Clin Invest. 2002; 110: 331–340.[CrossRef][Medline] [Order article via Infotrieve]
14. Detmers PA, Hernandez M, Mudgett J, Hassing H, Burton C, Mundt S, Chun S, Fletcher D, Card DJ, Lisnock J, Weikel R, Bergstrom JD, Shevell DE, Hermanowski-Vosatka A, Sparrow CP, Chao YS, Rader DJ, Wright SD, Pure E. Deficiency in inducible nitric oxide synthase results in reduced atherosclerosis in apolipoprotein E-deficient mice. J Immunol. 2000; 165: 3430–3435.
15. Chyu KY, Dimayuga P, Zhu J, Nilsson J, Kaul S, Shah PK, Cercek B. Decreased neointimal thickening after arterial wall injury in inducible nitric oxide synthase knockout mice. Circ Res. 1999; 85: 1192–1198.
16. Pfeifer A, Klatt P, Massberg S, Ny L, Sausbier M, Hirneiss C, Wang GX, Korth M, Aszodi A, Andersson KE, Krombach F, Mayerhofer A, Ruth P, Fassler R, Hofmann F. Defective smooth muscle regulation in cGMP kinase I-deficient mice. Embo J. 1998; 17: 3045–3051.[CrossRef][Medline] [Order article via Infotrieve]
17. Lincoln TM, Dey N, Sellak H. Invited review: cGMP-dependent protein kinase signaling mechanisms in smooth muscle: from the regulation of tone to gene expression. J Appl Physiol. 2001; 91: 1421–1430.
18. Feil R, Feil S, Hofmann F. A heretical view on the role of NO and cGMP in vascular proliferative diseases. Trends Mol Med. 2005; 11: 71–75.[CrossRef][Medline] [Order article via Infotrieve]
19. Garg UC, Hassid A. Nitric oxide-generating vasodilators and 8-bromo-cyclic guanosine monophosphate inhibit mitogenesis and proliferation of cultured rat vascular smooth muscle cells. J Clin Invest. 1989; 83: 1774–1777.[Medline] [Order article via Infotrieve]
20. Chiche JD, Schlutsmeyer SM, Bloch DB, de la Monte SM, Roberts JD, Jr, Filippov G, Janssens SP, Rosenzweig A, Bloch KD. Adenovirus-mediated gene transfer of cGMP-dependent protein kinase increases the sensitivity of cultured vascular smooth muscle cells to the antiproliferative and pro-apoptotic effects of nitric oxide/cGMP. J Biol Chem. 1998; 273: 34263–34271.
21. Boerth NJ, Dey NB, Cornwell TL, Lincoln TM. Cyclic GMP-dependent protein kinase regulates vascular smooth muscle cell phenotype. J Vasc Res. 1997; 34: 245–259.[Medline] [Order article via Infotrieve]
22. Anderson PG, Boerth NJ, Liu M, McNamara DB, Cornwell TL, Lincoln TM. Cyclic GMP-dependent protein kinase expression in coronary arterial smooth muscle in response to balloon catheter injury. Arterioscler Thromb Vasc Biol. 2000; 20: 2192–2197.
23. Sinnaeve P, Chiche JD, Gillijns H, Van Pelt N, Wirthlin D, Van De Werf F, Collen D, Bloch KD, Janssens S. Overexpression of a constitutively active protein kinase G mutant reduces neointima formation and in-stent restenosis. Circulation. 2002; 105: 2911–2916.
24. Takimoto E, Champion HC, Li M, Belardi D, Ren S, Rodriguez ER, Bedja D, Gabrielson KL, Wang Y, Kass DA. Chronic inhibition of cyclic GMP phosphodiesterase 5A prevents and reverses cardiac hypertrophy. Nat Med. 2005; 11: 214–222.[CrossRef][Medline] [Order article via Infotrieve]
25. Zhao L, Mason NA, Morrell NW, Kojonazarov B, Sadykov A, Maripov A, Mirrakhimov MM, Aldashev A, Wilkins MR. Sildenafil inhibits hypoxia-induced pulmonary hypertension. Circulation. 2001; 104: 424–428.
26. Mendelsohn ME. Viagra: now mending hearts. Nat Med. 2005; 11: 115–116.[CrossRef][Medline] [Order article via Infotrieve]
27. Wolfsgruber W, Feil S, Brummer S, Kuppinger O, Hofmann F, Feil R. A proatherogenic role for cGMP-dependent protein kinase in vascular smooth muscle cells. Proc Natl Acad Sci USA. 2003; 100: 13519–13524.
28. Kumar A, Lindner V. Remodeling with neointima formation in the mouse carotid artery after cessation of blood flow. Arterioscler Thromb Vasc Biol. 1997; 17: 2238–2244.
29. Lindner V, Fingerle J, Reidy MA. Mouse model of arterial injury. Circ Res. 1993; 73: 792–796.
30. Holtwick R, Gotthardt M, Skryabin B, Steinmetz M, Potthast R, Zetsche B, Hammer RE, Herz J, Kuhn M. Smooth muscle-selective deletion of guanylyl cyclase-A prevents the acute but not chronic effects of ANP on blood pressure. Proc Natl Acad Sci USA. 2002; 99: 7142–7147.
31. Wegener JW, Nawrath H, Wolfsgruber W, Kuhbandner S, Werner C, Hofmann F, Feil R. cGMP-dependent protein kinase I mediates the negative inotropic effect of cGMP in the murine myocardium. Circ Res. 2002; 90: 18–20.
32. Soriano P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet. 1999; 21: 70–71.[CrossRef][Medline] [Order article via Infotrieve]
33. Li L, Miano JM, Cserjesi P, Olson EN. SM22 alpha, a marker of adult smooth muscle, is expressed in multiple myogenic lineages during embryogenesis. Circ Res. 1996; 78: 188–195.
34. Geiselhoringer A, Gaisa M, Hofmann F, Schlossmann J. Distribution of IRAG and cGKI-isoforms in murine tissues. FEBS Lett. 2004; 575: 19–22.[CrossRef][Medline] [Order article via Infotrieve]
35. Yogo K, Shimokawa H, Funakoshi H, Kandabashi T, Miyata K, Okamoto S, Egashira K, Huang P, Akaike T, Takeshita A. Different vasculoprotective roles of NO synthase isoforms in vascular lesion formation in mice. Arterioscler Thromb Vasc Biol. 2000; 20: e96–e100.
36. Monks D, Lange V, Silber RE, Markert T, Walter U, Nehls V. Expression of cGMP-dependent protein kinase I and its substrate VASP in neointimal cells of the injured rat carotid artery. Eur J Clin Invest. 1998; 28: 416–423.[CrossRef][Medline] [Order article via Infotrieve]
37. Hanafy KA, Krumenacker JS, Murad F. NO, nitrotyrosine, and cyclic GMP in signal transduction. Med Sci Monit. 2001; 7: 801–819.[Medline] [Order article via Infotrieve]
38. Foster MW, McMahon TJ, Stamler JS. S-nitrosylation in health and disease. Trends Mol Med. 2003; 9: 160–168.[CrossRef][Medline] [Order article via Infotrieve]
39. Ying J, Tong X, Pimentel DR, Weisbrod RM, Trucillo MP, Adachi T, Cohen RA. Cysteine-674 of the sarco/endoplasmic reticulum calcium ATPase is required for the inhibition of cell migration by nitric oxide. Arterioscler Thromb Vasc Biol. 2007; 27: 783–790.
40. Tulis DA, Bohl Masters KS, Lipke EA, Schiesser RL, Evans AJ, Peyton KJ, Durante W, West JL, Schafer AI. YC-1-mediated vascular protection through inhibition of smooth muscle cell proliferation and platelet function. Biochem Biophys Res Commun. 2002; 291: 1014–1021.[CrossRef][Medline] [Order article via Infotrieve]
41. Sinnaeve P, Chiche JD, Nong Z, Varenne O, Van Pelt N, Gillijns H, Collen D, Bloch KD, Janssens S. Soluble guanylate cyclase alpha(1) and beta(1) gene transfer increases NO responsiveness and reduces neointima formation after balloon injury in rats via antiproliferative and antimigratory effects. Circ Res. 2001; 88: 103–109.
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